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A pilot study of non-targeted screening for stimulant misuse using high-resolution mass spectrometry

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Abstract

Purpose

Forensic and anti-doping laboratories have used strategies based on targeted approaches in data processing, which means that only targeted compounds have been analyzed. The new challenges lie beyond the anticipation of molecular targets, such as the detection of designer drugs. In this research, after automatic extraction of all signals obtained by liquid chromatography–high-resolution mass spectrometry (LC–HRMS), the atypical pattern of stimulant-positive samples is evaluated to indicate potential doping substances by principal component analysis (PCA) when compared to negative samples.

Methods

The capacity for discrimination of positive samples by PCA was evaluated using 24 samples reported as an adverse analytical finding (AAF) for stimulants from different sports against their corresponding negative controls. The method was then evaluated on routine analysis of 620 samples.

Results

AAF samples for stimulants presented an atypical pattern positioning when compared to negative samples in the multidimensional space of PCA. The method was able to identify 22 samples reported as AAF before checking the targeted substance windows. In that scenario, it also allows the detection of designer stimulants when the present procedure is applied complementarily in screening routine analysis. The present approach was applied simultaneously with the targeted procedure through the evaluation of 620 samples without jeopardizing the results deadline.

Conclusions

Non-targeted analysis of stimulants by PCA as applied to athlete urine samples was demonstrated for the first time to be capable of flagging positive samples in sport competitions, showing its promise as a complementary screening tool alongside classical methods and as a straightforward approach for selecting samples for long-term storage.

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References

  1. Pereira HMG, Sardela VF, Padilha MC, Mirotti L, Casilli A, de Oliveira FA, de Albuquerque Cavalcanti G, Rodrigues LML, de Araujo ALD, Levy RS, Teixeira PAC, de Oliveira FAG, Duarte ACG, Carneiro ACD, Evaristo JAM, Dos Santos GRC, da Costa GCV, de Lima Castro F, Nogueira FCS, Scalco FB, Pizzatti L, de Aquino Neto FR (2017) Doping control analysis at the Rio 2016 Olympic and Paralympic games. Drug Test Anal 9:1658–1672

    Article  CAS  PubMed  Google Scholar 

  2. Sobolevsky T, Krotov G, Dikunets M, Nikitina M, Mochalova E, Rodchenkov G (2014) Anti-doping analyses at the Sochi Olympic and Paralympic Games 2014. Drug Test Anal 6:1087–1101

    Article  CAS  PubMed  Google Scholar 

  3. de Aquino Neto FR, Sarderla VF, Mirotti L, Pizzatti L (2016) Running ahead of doping: analytical advances and challenges faced by modern laboratories ahead of Rio 2016. Bioanalysis 8:1753–1756

    Article  CAS  PubMed  Google Scholar 

  4. World Anti-Doping Agency (2016) International standard for laboratories (ISL). Version 9.0, WADA, Montreal. https://www.wada-ama.org/sites/default/files/resources/files/isl_june_2016.pdf. Accessed Dec 2018

  5. McLaren RH (2016) Richard H. McLaren independent person, WADA investigation of Sochi allegations. WADA, Montreal. https://www.wada-ama.org/sites/default/files/resources/files/20160718_ip_report_newfinal.pdf. Accessed Dec 2018

  6. Ellis DI, Dunn WB, Griffin JL, Allwood JW, Goodacre R (2007) Metabolic fingerprinting as a diagnostic tool. Pharmacogenomics 8:1243–1266

    Article  CAS  PubMed  Google Scholar 

  7. Kaddurah-Daouk R, Weinshilboum R (2015) Metabolomic signatures for drug response phenotypes: pharmacometabolomics enables precision medicine. Clin Pharmacol Ther 98:71–75

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Kiss A, Lucio M, Fildier A, Buisson C, Schmitt-Kopplin P, Cren-Olivé C (2013) Doping control using high and ultra-high-resolution mass spectrometry based non-targeted metabolomics—a case study of salbutamol and budesonide abuse. PLoS One 8:e74584. https://doi.org/10.1371/journal.pone.0074584

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Raro M, Ibáñez M, Gil R, Fabregat A, Tudela E, Deventer K, Ventura R, Segura J, Marcos J, Kotronoulas A, Joglar J, Farré M, Yang S, Xing Y, Van Eenoo P, Pitarch E, Hernández F, Sancho JV, Pozo ÓJ (2015) Untargeted metabolomics in doping control: detection of new markers of testosterone misuse by ultrahigh performance liquid chromatography coupled to high-resolution mass spectrometry. Anal Chem 87:8373–8380

    Article  CAS  PubMed  Google Scholar 

  10. Kiss A, Jacquet A-L, Paisse O, Flament-Waton M-M, de Ceaurriz J, Bordes C, Gauvrit J-Y, Lantéri P, Cren-Olivé C (2011) Urinary signature of anabolic steroids and glucocorticoids in humans by LC–MS. Talanta 83:1769–1773

    Article  CAS  PubMed  Google Scholar 

  11. Alladio E, Caruso R, Gerace E, Amante E, Salomone A, Vincenti M (2016) Application of multivariate statistics to the steroidal module of the athlete biological passport: a proof of concept study. Anal Chim Acta 922:19–29

    Article  CAS  PubMed  Google Scholar 

  12. Sardela VF, Martucci MEP, de Araújo ALD, Leal EC, Oliveira DS, Carneiro GRA, Deventer K, Van Eenoo P, Pereira HMG, Aquino Neto FR (2018) Comprehensive analysis by liquid chromatography Q-Orbitrap mass spectrometry: fast screening of peptides and organic molecules. J Mass Spectrom 53:476–503

    Article  CAS  PubMed  Google Scholar 

  13. Cawley A, Pasin D, Ganbat N, Ennis L, Smart C, Greer C, Keledjian J, Fu S, Chen A (2016) The potential for complementary targeted/non-targeted screening of novel psychoactive substances in equine urine using liquid chromatography-high resolution accurate mass spectrometry. Anal Methods 8:1789–1797

    Article  CAS  Google Scholar 

  14. Saugy M, Robinson N, Saudan C (2009) The fight against doping: back on track with blood. Drug Test Anal 1:474–478

    Article  CAS  PubMed  Google Scholar 

  15. Ransohoff DF (2005) Bias as a threat to the validity of cancer molecular-marker research. Nat Rev Cancer 5:142–149

    Article  CAS  PubMed  Google Scholar 

  16. Zhang S, Liu L, Steffen D, Ye T, Raftery D (2012) Metabolic profiling of gender: headspace-SPME/GC–MS and 1H NMR analysis of urine. Metabolomics 8:323–334

    Article  CAS  Google Scholar 

  17. Yan B, A J, Wang G, Lu H, Huang X, Liu Y, Zha W, Hao H, Zhang Y, Liu L, Gu S, Huang Q, Zheng Y, Sun J (2009) Metabolomic investigation into variation of endogenous metabolites in professional athletes subject to strength-endurance training. J Appl Physiol 106:531–538

    Article  CAS  PubMed  Google Scholar 

  18. Al-Khelaifi F, Diboun I, Donati F, Botrè F, Alsayrafi M, Georgakopoulos C, Suhre K, Yousri NA, Elrayess MA (2018) A pilot study comparing the metabolic profiles of elite-level athletes from different sporting disciplines. Sports Med Open 4:2. https://doi.org/10.1186/s40798-017-0114-z

    Article  PubMed  PubMed Central  Google Scholar 

  19. Janssen PAJ, Leysen JE, Megens AAHP, Awouters FHL (1999) Does phenylethylamine act as an endogenous amphetamine in some patients? Int J Neuropsychopharmacol 2:229–240

    Article  CAS  PubMed  Google Scholar 

  20. Cone EJ, Tsadik A, Oyler J, Darwin WD (1998) Cocaine metabolism and urinary excretion after different routes of administration. Ther Drug Monit 20:556–560

    Google Scholar 

  21. Current JD (2004) Pharmacology for anesthetists. PediaPress, Mainz

    Article  CAS  PubMed  Google Scholar 

  22. World Anti-Doping Agency (2010) The 2010 prohibited list international standard. WADA, Montreal. https://www.wada-ama.org/sites/default/files/resources/files/WADA_Prohibited_List_2010_EN.pdf. Accessed Dec 2018

  23. Pereira HMG, Sardela VF (2014) Stimulant doping agents used in Brazil: prevalence, detectability, analytical implications, and challenges. Subst Use Misuse 49:1098–1114

    Article  PubMed  Google Scholar 

  24. Perrenoud L, Saugy M, Saudan C (2009) Detection in urine of 4-methyl-2-hexaneamine, a doping agent. J Chromatogr B 877:3767–3770

    Article  CAS  Google Scholar 

  25. Deventer K, Roels K, Delbeke FT, Van Eenoo P (2011) Prevalence of legal and illegal stimulating agents in sports. Anal Bioanal Chem 401:421–432

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We wish to thank Thermo Fisher Scientific and Nova Analítica for providing technical assistance for this project. The technical assistance of Igor Rodrigues da Costa and Professor Rafael Garrett da Costa is also greatly appreciated. This work was financed in part/collaterally by the following Brazilian research-sponsoring foundations: CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), FAPERJ (Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior–Brasil (CAPES)–Finance Code 001).

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Authors and Affiliations

  1. Universidade Federal do Rio de Janeiro, Instituto de Química, LBCD-LADETEC, Avenida Horácio Macedo, nº 1281, Polo de Química, bloco C, Cidade Universitária, Rio de Janeiro, RJ, 21941-598, Brazil

    Patrícia Davies de Oliveira Sardela, Vinícius Figueiredo Sardela, Andressa Maia dos Santos da Silva, Henrique Marcelo Gualberto Pereira & Francisco Radler de Aquino Neto

  2. Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Campus São Gonçalo, Rua José Augusto Pereira dos Santos s/nº, São Gonçalo, RJ, 24425-004, Brazil

    Patrícia Davies de Oliveira Sardela

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  1. Patrícia Davies de Oliveira Sardela
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  2. Vinícius Figueiredo Sardela
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  3. Andressa Maia dos Santos da Silva
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  4. Henrique Marcelo Gualberto Pereira
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  5. Francisco Radler de Aquino Neto
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Corresponding author

Correspondence to Patrícia Davies de Oliveira Sardela.

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The authors declare that they have no conflict of interest.

Ethical approval

The urine samples were obtained from male athletes after getting their consent for research in the doping control form. This article does not contain any studies with animals performed by any of the authors.

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Sardela, P.D., Sardela, V.F., da Silva, A.M. et al. A pilot study of non-targeted screening for stimulant misuse using high-resolution mass spectrometry. Forensic Toxicol 37, 465–473 (2019). https://doi.org/10.1007/s11419-019-00482-1

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